scholarly journals Rapid Detection of Campylobacter coli, C. jejuni, and Salmonella enterica on Poultry Carcasses by Using PCR-Enzyme-Linked Immunosorbent Assay

2003 ◽  
Vol 69 (6) ◽  
pp. 3492-3499 ◽  
Author(s):  
Yang Hong ◽  
Mark E. Berrang ◽  
Tongrui Liu ◽  
Charles L. Hofacre ◽  
Susan Sanchez ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Salmonella Enterica ◽  
Rapid Detection ◽  
Diarrheal Disease ◽  
False Negative ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Coli ◽  
Culture Methods ◽  
Isolation And Identification ◽  
Serological Tests

ABSTRACT Contamination of retail poultry by Campylobacter spp. and Salmonella enterica is a significant source of human diarrheal disease. Isolation and identification of these microorganisms require a series of biochemical and serological tests. In this study, Campylobacter ceuE and Salmonella invA genes were used to design probes in PCR-enzyme-linked immunosorbent assay (ELISA), as an alternative to conventional bacteriological methodology, for the rapid detection of Campylobacter jejuni, Campylobacter coli, and S. enterica from poultry samples. With PCR-ELISA (40 cycles), the detection limits for Salmonella and Campylobacter were 2 � 102 and 4 � 101 CFU/ml, respectively. ELISA increased the sensitivity of the conventional PCR method by 100- to 1,000-fold. DNA was extracted from carcass rinses and tetrathionate enrichments and used in PCR-ELISA for the detection of Campylobacter and S. enterica, respectively. With PCR-ELISA, Salmonella was detected in 20 of 120 (17%) chicken carcass rinses examined, without the inclusion of an enrichment step. Significant correlation was observed between PCR-ELISA and cultural methods (kappa = 0.83; chi-square test, P < 0.001) with only one false negative (1.67%) and four false positives (6.67%) when PCR-ELISA was used to screen 60 tetrathionate enrichment cultures for Salmonella. With PCR-ELISA, we observed a positive correlation between the ELISA absorbance (optical density at 405 nm) and the campylobacter cell number in carcass rinse, as determined by standard culture methods. Overall, PCR-ELISA is a rapid and cost-effective approach for the detection and enumeration of Salmonella and Campylobacter bacteria on poultry.

scholarly journals Rapid Detection of Salmonella enterica Serovar Choleraesuis in Blood Cultures by a Dot Blot Enzyme-Linked Immunosorbent Assay

2000 ◽  
Vol 7 (6) ◽  
pp. 977-979 ◽  
Author(s):  
Kritsana Janyapoon ◽  
Sunee Korbsrisate ◽  
Hatairat Thamapa ◽  
Sittichai Thongmin ◽  
Suwattana Kanjanahareutai ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Salmonella Enterica ◽  
Rapid Detection ◽  
Direct Detection ◽  
Blood Cultures ◽  
Enzyme Linked Immunosorbent Assay ◽  
Biochemical Method ◽  
Dot Blot

ABSTRACT A dot blot enzyme-linked immunosorbent assay (ELISA) with a monoclonal antibody specific to phase1-c Salmonella was developed for the direct detection of Salmonella entericaserovar Choleraesuis in blood cultures. This system was applied to the identification of serovar Choleraesuis, and the results were compared with those obtained by a conventional biochemical method. It was revealed that all 12 samples identified to be infected with serovar Choleraesuis were positive on testing by the ELISA. In contrast, 77 samples infected with bacteria commonly isolated from the blood were not reactive by the ELISA. The calculated sensitivity and specificity of the established assay are 100%.

An Enzyme-Linked Immunosorbent Assay for the Rapid Detection of Staphylococcus aureus in Processed Foods

1994 ◽  
Vol 57 (3) ◽  
pp. 184-189 ◽  
Author(s):  
TSUNG C. CHANG ◽  
SU H. HUANG
Keyword(s):  
Staphylococcus Aureus ◽  
Immunosorbent Assay ◽  
Rapid Detection ◽  
Protein A ◽  
Large Degree ◽  
Microtiter Plate ◽  
Specific Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Processed Foods ◽  
Culture Methods

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the rapid detection of Staphylococcus aureus in foods. The assay was based on the detection of protein A which is a specific protein secreted by S. aureus. Following a 24-h incubation in a staphylococcal selective broth containing mannitol as the carbon source, the culture supematant was added to the microtiter plate coated with anti-protein A immunoglobulin G (IgG). After incubation, peroxidase-labeled anti-protein A IgG was used to produce the signal of antigen-antibody reaction. The sensitivity of the assay for protein A was 0.1 ng/ml. For 37 strains of S. aureus studied, all produced protein A, and the amount (13-1,100 ng/ml) of protein A secreted by different strains varied to a large degree. For another 57 strains (including 19 Staphylococcus spp.) of bacteria tested, two strains (5. capilis subsp. capitis CCRC 12161 and S. lentus CCRC 12926) produced very low amounts of protein A (0.6-1 ng/ml) after 24-h incubation. Staphylococcus aureus was detected by the ELISA in all of six samples of precooked foods naturally contaminated with the bacterium. Twenty-two processed foods artificially inoculated with S. aureus at levels of &lt; 2 CFU/g and 10 to 20 CFU/g, respectively, were all positive by the ELISA. As compared to the conventional culture methods which take 5 to 6 days to complete, the ELISA can detect low numbers of S. aureus in processed foods with a total analytical time of only 28 h.

scholarly journals Sensitivity, Specificity, and Predictive Values of ThreeSalmonella Rapid Detection Kits Using Fresh and Frozen Poultry Environmental Samples versus Those of Standard Plating

1999 ◽  
Vol 65 (3) ◽  
pp. 1055-1060 ◽  
Author(s):  
Melissa O. Peplow ◽  
Maria Correa-Prisant ◽  
Martha E. Stebbins ◽  
Frank Jones ◽  
Peter Davies
Keyword(s):  
Rapid Detection ◽  
Environmental Samples ◽  
False Negative ◽  
Food Samples ◽  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Culture Methods ◽  
Predictive Values ◽  
Negative Results ◽  
Rapid Tests

ABSTRACT To reduce human exposure to Salmonella spp. in poultry products, broiler chicken flocks have been tested by culture methods. Since the standard techniques may take 3 to 5 days, rapid detection methods have been developed. In this study we tested the performance of three rapid tests originally developed for food samples by using environmental samples obtained from poultry houses. These rapid tests were Reveal, an enzyme-linked immunosorbent assay from Neogen Corp.; BIND, a bacterial ice nucleation detection method from Idetek Corp.; and a filter monitor method from Future Medical Technologies, Inc. For the standard culture, brilliant green with novabiocin and xylose–lysine–tergitol-4 agar were used for presumptive identification, and identities were confirmed by using poly-O antisera. Environmental samples were collected from farms belonging to an integrated poultry company prior to chick placement and 1 week before slaughter. Sensitivities, specificities, and predictive values with 95% confidence intervals were calculated. Statistical differences were determined by using McNemar’s chi square test. The sensitivities of the different tests were not stable, varying widely between sample times, and were affected by freezing of the samples. All of the rapid tests had low sensitivities, which led to many false-negative results. All tests were able to detect Salmonella spp. at a concentration of 10 CFU/ml in at least one of four trials. The BIND and Reveal tests were simple to use with multiple samples and reduced laboratory time by up to 1 day. Based on our results, we do not recommend that any of these rapid tests, in their present state of development, be utilized with environmental samples collected with drag swabs.

Rapid detection of human rotavirus strains in stools by single-sandwich enzyme-linked immunosorbent assay systems using monoclonal antibodies

1989 ◽  
Vol 24 (1-2) ◽  
pp. 43-56 ◽  
Author(s):  
Giuseppe Gerna ◽  
Antonella Sarasini ◽  
Angela Di Matteo ◽  
Maurizio Parea ◽  
Maria Torseilini ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Rapid Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Rotavirus

Secretory Anti-Giardia lamblia Antibodies in Human Milk: Protective Effect Against Diarrhea

PEDIATRICS ◽  
1994 ◽  
Vol 93 (1) ◽  
pp. 28-31
Author(s):  
Juan N. Walterspiel ◽  
Ardythe L. Morrow ◽  
Larry K. Pickering ◽  
Guillermo M. Ruiz-Palacios ◽  
M. Lourdes Guerrero
Keyword(s):  
Human Milk ◽  
Protective Effect ◽  
Immunosorbent Assay ◽  
Giardia Lamblia ◽  
Diarrheal Disease ◽  
Secretory Iga ◽  
Enzyme Linked Immunosorbent Assay ◽  
Giardia Infection ◽  
Maternal Milk ◽  
Milk Samples

Objective. To determine whether anti-Giardia lamblia secretory IgA (sIgA) antibodies in human milk protect infants from acquisition of or symptoms associated with Giardia infection. Methods. One hundred ninety-seven Mexican mother/infant pairs were followed weekly from birth for diarrheal disease and feeding status. Infant stool specimens were collected weekly and were cultured for bacterial pathogens and tested for Giardia and rotavirus by enzyme-linked immunosorbent assay. Maternal milk samples were collected weekly for 1 month postpartum and monthly thereafter. To determine the protective effect of anti-Giardia sIgA in milk against infection and against diarrhea due to Giardia, milk samples from mothers of infected infants and appropriately matched controls were assayed for anti-Giardia sIgA by enzyme-linked immunosorbent assay. Results. Asymptomatic, infected infants ingested significantly (P = .046) higher amounts of milk anti-Giardia sIgA compared with symptomatic, infected infants. However, milk anti-Giardia sIgA concentrations did not differ between Giardia-infected and noninfected infants. Conclusion. The amount of anti-Giardia sIgA in human milk was associated with prevention of symptoms of diarrhea due to Giardia, but not with acquisition of the organism.

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